Highly effective detection of human papillomavirus 16 and 18 DNA by a testing algorithm combining broad-spectrum and type-specific PCR

J Clin Microbiol. 2006 Sep;44(9):3292-8. doi: 10.1128/JCM.00539-06.

Abstract

The use of a single broad-spectrum human papillomavirus (HPV) DNA-based PCR test may fail to detect lower concentrations of HPV DNA due to competition between different genotypes in mixed infections. To improve HPV detection by PCR, broad-spectrum and type-specific (TS) PCRs were combined, with a focus on HPV-16 and HPV-18. Cervical and cervicovaginal cell samples were obtained from 1,113 healthy women (age range, 15 to 25 years) participating in an HPV-16/HPV-18 candidate vaccine efficacy trial. These samples were tested by a broad-spectrum SPF(10) PCR-DNA enzyme immunoassay, followed by a primer SPF(10)-based line probe assay (SPF(10) LiPA), and HPV-16- and HPV-18-TS PCRs. The results for the majority of the HPV-16/18 SPF(10) LiPA-positive samples were confirmed by TS-PCR (kappa values, 0.775 for HPV-16 and 0.785 for HPV-18). However, TS PCR revealed additional positive samples among those that contained other HPV genotypes due to competition. Conversely, SPF(10) LiPA identified HPV-16 or -18 in samples that remained negative by TS PCR as a result of sampling variation. Analysis of follow-up samples from more than 1,000 women confirmed that the combination of SPF(10)-LiPA with additional HPV-16- and HPV-18-TS PCR diminishes the rate of false-negative diagnosis. The combination of broad-spectrum and TS PCRs resulted in a novel testing algorithm. This combination of assays is more accurate than either method alone, and the novel algorithm offers a highly accurate and effective method for the analysis of HPV infections.

Publication types

  • Clinical Trial, Phase II
  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Algorithms*
  • Cervix Uteri / virology
  • DNA Probes, HPV
  • DNA, Viral / analysis*
  • DNA, Viral / isolation & purification
  • Female
  • Human papillomavirus 16 / classification
  • Human papillomavirus 16 / genetics
  • Human papillomavirus 16 / isolation & purification*
  • Human papillomavirus 18 / classification
  • Human papillomavirus 18 / genetics
  • Human papillomavirus 18 / isolation & purification*
  • Humans
  • Papillomavirus Infections / diagnosis*
  • Papillomavirus Infections / prevention & control
  • Papillomavirus Infections / virology
  • Polymerase Chain Reaction / methods*
  • Species Specificity
  • Treatment Outcome
  • Viral Vaccines / therapeutic use

Substances

  • DNA Probes, HPV
  • DNA, Viral
  • Viral Vaccines